蛋白激酶C在糖基化终末产物介导结肠平滑肌细胞内钙离子浓度中的作用  

Advanced glycation end products inhibit intracellular calcium concentration in colon smooth muscle cells in a protein kinase C-dependent manner

作　　者：朱滢[1] 王庆娥[1] 王云[1] 巩尧瑶[1] 孙晓萌[1] 林琳[1] 

机构地区：[1]南京医科大学第一附属医院消化科,江苏省南京市210029

出　　处：《世界华人消化杂志》2014年第6期874-879,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81270462;江苏省国际合作基金资助项目;No.BZ2011044~~

摘　　要：目的:探讨糖基化终末产物(advanced glycation end products,AGEs)对结肠平滑肌细胞内钙离子浓度的影响及可能机制.方法:酶解法分离培养SD大鼠结肠平滑肌细胞(smooth muscle cell,SMC),α-actin免疫荧光鉴定;激光共聚焦显微镜检测(SMC)钙闪烁;蛋白激酶C(protein kinase C,PKC)活性检测试剂盒检测细胞PKC活性.结果:细胞免疫荧光鉴定大鼠结肠SMC.不同浓度AGEs(50、100μg/mL)刺激结肠SMC后,与对照组比较,钙离子浓度显著降低(56.7%±3.6%、78.6%±5%vs 99.6%±3.1%,P<0.05,P<0.01),50μg/mL为体外最大有效浓度.与对照组相比,AGEs(50、100μg/mL)升高细胞内PKC活性.PKC抑制剂chelerythrine可阻断A G E s介导的钙离子浓度降低(70.7%±3.7%vs 87.1%±2.5%,P<0.05).结论:AGEs可激活PKC通路、从而降低胞内钙离子的浓度,最终抑制大鼠结肠平滑肌收缩.AIM： To investigate the effect of advanced gly- cation end products （AGEs） on intracellular calcium concentration in isolated colonic smooth muscle cells and the possible mechanisms in- volved.isolated from normal adult rats, and immumo- fluorescence staining for α-actin was used to identify smooth muscle cells. The responsive- ness of colonic smooth muscle cells to AGEs was measured by confocal laser scanning microscopy. Intracellular Ca2＋ concentration （[Ca2＋]i） was determined by Fluo3/AM based digital microfluorimetric measurement. Protein kinase C （PKC） activity was detected by PKC activity assay. PKC inhibitor chelerythrine was used to examine the role of PKC in AGEs- mediated inhibition of [Ca2±]i in colonic smooth muscle cells. RESULTS： Colonic smooth muscle cells were successfully isolated from normal rats and iden- tified by immunofluorescence staining. AGEs inhibited [Ca2＋]i in a concentration-dependent manner. AGEs at a concentration of 50 or 100 μg/mL significantly inhibited the mean [Ca2＋]i compared with the control group （56.7% ± 3.6%, 78.6% ~ 5% vs 99.6% ± 3.1%, P 〈 0.05, P 〈 0.01）. PKC activity increased in SMCs treated with 50 μg/mL or 100 μg/mL of AGEs compared with the control group. Pretreatment with chelery- thrine （1 μmol/L） reduced AGEs-mediated inhi- bition of [Ca2＋]i （70.7% ± 3.7% vs 87.1% ± 2.5%, P 〈 0.05） CONCLUSION： AGEs inhibit [Ca2＋]i in colonic smooth muscle cells in a PKC-dependent man- ner.

关 键 词：糖基化终末产物 结肠平滑肌细胞 蛋白激 酶c 钙离子 

分 类 号：R587.2[医药卫生—内分泌]